pRS305 (ATCC® 77140)

Applications: YI-type (integrating) shuttle vectorshuttle vectorvector containing primer sites useful for sequencingvector permitting RNA synthesis in vitrovector permitting production of single-stranded DNAvector permitting visual detection of recombinants  /  Depositors: P Hieter

Designations pRS305
Depositors P Hieter
Biosafety Level 1
Vector Information
Size (kb): 5.5040001869201660
Vector: pRS305 (phagemid)
Promoters: Promoter for in vitro transcription T7
Construction: pRSS56 [pBluescript KS+, pBS(+)]
Marker(s):LEU2,ampR
Construct size (kb): 5.504000186920166
Features: insert detection: lacZ'
marker(s): LEU2
marker(s): ampR
promoter: lac
promoter for in vitro transcription: T3
promoter for in vitro transcription: T7
replicon: f1
replicon: pMB1
MCS: SacI...KpnI
Applications
YI-type (integrating) shuttle vector
shuttle vector
vector containing primer sites useful for sequencing
vector permitting RNA synthesis in vitro
vector permitting production of single-stranded DNA
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): HindIII--5.6; PvuII--5.1, 0.46; KpnI--3.8, 1.7; EcoRI--3.5, 2.0.
One of a series of pBluescript-based integrating vectors (ATCC 77138-77141) differing in the yeast selectable marker gene.
YI-type integrating shuttle vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and encodes the lacZ alpha (lacZ') peptide.
pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites between bla and f1 origin of pBS(+).
A fragment (2.235 kb) containing the LEU2 gene was inserted into the NdeI site of pRSS56. All ends were blunted.
The order of the major features in this plasmid is: LEU2 - f1 ori (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla.
The following restriction sites in the multiple cloning site (MCS) are no longer unique: KpnI ClaI EcoRV EcoRI BstXI.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122: 19-27, 1989. PubMed: 2659436

Related Products
component of:ATCC 77189
Shipped frozen
Shipping Information Freeze dried phagemid in E. coli

Product Sheet
Product Sheet
Product Sheet
Attachments Vector map for 77140